Activity assays of nine heterogeneous promoters in neural and other cultured cells

In Vitro Cell Dev Biol Anim. 1994 May;30A(5):300-5. doi: 10.1007/BF02631450.

Abstract

To express high levels of proteins encoded by transfected DNA constructs in a variety of cultured cells, including neuronal cells, the activities of nine different promoters were evaluated using Escherichia coli beta-galactosidase (beta-gal) (LacZ) as a reporter gene. These nine promoters were categorized into three distinct groups (high, intermediate, and low expresser), in terms of the levels of beta-gal expression. An expression vector containing the cytomegalovirus enhancer and the chick beta-actin promoter (high expresser) showed the highest levels of expression, followed by vectors containing the cytomegalovirus promoter/enhancer and the SV40 promoter/enhancer (intermediate expresser). The rest of the promoters (thymidine kinase, adenovirus, murine proliferative sarcoma virus, nerve growth factor receptor, Rous sarcoma and mouse mammary tumor virus, and beta-amyloid precursor protein) expressed low levels of beta-gal. These results were consistent for eight different cell types. A particularly attractive model is the stem cell, P19; cultures differentiating into progeny consisting predominantly of cholinergic neurons could be readily transfected with expression vectors using liposomes and expressed beta-gal without significant morphologic changes of the differentiated neurons. The systems should be useful for the study of promoters and various expressed proteins, including those involved in axonal transport.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • Cell Line
  • Chlorocebus aethiops
  • Enhancer Elements, Genetic
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Evaluation Studies as Topic
  • Genes, Reporter
  • Genetic Vectors
  • Humans
  • Mice
  • Neurons / cytology
  • Neurons / metabolism*
  • Promoter Regions, Genetic*
  • Transfection
  • Tretinoin / pharmacology
  • Tumor Cells, Cultured
  • beta-Galactosidase / genetics

Substances

  • Tretinoin
  • beta-Galactosidase